ゼラチンとリン酸カルシウムで表面修飾したPET 上でのMC3T3-E1 細胞の増殖と石灰化

抄録

With the aim of developing a new type of non-resorbable membrane for guided bone regeneration (GBR) that could promote bone formation, a surface-modified poly(ethyleneterephthalate)(PET) sheet coated with a double layer of gelatin and calcium phosphate was prepared and the response of MC3T3-E1 cells was investigated. The material was prepared through the following steps: surface hydrolysis of PET sheet, covalent bonding of a gelatin layer, immobilization of urease, and deposition of calcium phosphate layer with the mediation of urease at 310 K. The gelatin layer was very thin and the calcium phosphate layer was 5 to 10 μm thick. The calcium phosphate layer consisted of octacalcium phosphate (OCP) and low-crystallinity hydroxyapatite (HA) , and dissolved in saline up to the saturation concentration within one day. MC3T3-E1 cells were cultured on untreated PET, PET coated with gelatin (PET/gel), and PET coated with gelatin and calcium phosphate (PET/gel/CaP) for up to 60 days. Although the cells did not attach on untreated PET, they attached and proliferated on PET/gel. The proliferation was more pronounced on PET/gel/CaP. In addition, deposition of calcium salts was evidenced by von Kossa staining on PET/gel/CaP. The calcium phosphate layer was considered to act as one of the supply sources of calcium ions to assist the differentiation of MC3T3-E1, and the gelatin layer may have acted as the platform for the attachment and proliferation of the cells. The present material is therefore considered to be a promising candidate for a non-resorbable GBR membrane that could assist bone formation.